Intein technology to deliver ABCA3 lipid transporter for ABCA3 deficiency
Splitting ABCA3 cDNA into fragments using intein technology for AAV delivery addresses the inefficiencies in ABCA3 deficiency treatment, enabling effective protein recombination and potential therapeutic benefits.
US20250235552A1Pending Publication Date: 2025-07-24CHILDRENS HOSPITAL MEDICAL CENT CINCINNATI
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Patent Information
- Application Number
- US18/415949
- Authority / Receiving Office
- US · United States
- Patent Type
- Applications(United States)
- Current Assignee / Owner
- Filing Date
- 2024-01-18
- Publication Date
- 2025-07-24
AI Technical Summary
Technical Problem
Existing methods to treat ABCA3 deficiency caused by splicing mutations are inefficient, and full-length ABCA3 cDNA cannot be packaged into AAV viruses due to its large size, posing a challenge for effective therapeutic delivery.
Method used
Splitting ABCA3 cDNA into two fragments using intein technology for separate AAV delivery, allowing recombination into full-length ABCA3 protein in lung alveolar type II cells, and using CRISPR/Cas9 to delete a loxP site in ABCA3 flox mice.
Benefits of technology
Successfully produced recombined ABCA3 protein in vitro and plan to assess its therapeutic potential in ABCA3 deficient mice, potentially extending survival rates.
✦ Generated by Eureka AI based on patent content.
Abstract
In order to develop a therapeutic strategy to rescue ABCA3 deficiency caused by a splicing mutation of ABCA3, we have designed an experimental scheme using CRISPR / Cas9 to delete a loxP site located at an intronic region of ABCA3 flox mice (Sftpc-Cre;Abca3flox / flox). The deletion of this loxP site extended survival of the mice by only a week, probably due to insufficient re-ligation efficiency after the CRISPR / Cas9 deletion. An alternative approach to rescuing ABCA3 deficiency caused by mutations in ABCA3, including coding or non-coding (e.g., intronic) mutations, is to deliver intact ABCA3 into lung alveolar type II cells using AAV (e.g., AAV6.2FF), a clinically approved delivery system. Due to the large size of ABCA3 (1704 amino acids), it is technically impossible to package full length ABCA3 cDNA into an AAV virus. However, a recent technological advance using intein makes it possible to split ABCA3 cDNA into two fragments for separate AAV delivery to the cells wherein the translated ABCA3 protein fragments would recombine into full length ABCA3 protein.
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